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Ripa buffer

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RIPA buffer is a commonly used cell lysis and protein extraction buffer. It is a solution designed to solubilize proteins from cells or tissues for subsequent analysis. The buffer contains a combination of detergents, salts, and other components that help to disrupt cell membranes and release proteins into the solution.

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Lab products found in correlation

Mab casper 1, by Adipogen (1 mentions) Horseradish peroxidase conjugated anti mouse and anti rabbit igg, by Cell Signaling Technology (1 mentions) Nupage, by Thermo Fisher Scientific (1 mentions) Bicinchoninic acid (bca), by Thermo Fisher Scientific (1 mentions) Protease inhibitor, by Merck Group (1 mentions) Protease inhibitor, by Thermo Fisher Scientific (1 mentions) Phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)

2 protocols using ripa buffer

1

Inflammasome Protein Detection in Cells

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Cells were lysed in RIPA buffer (R&D Systems) containing protease inhibitors (Sigma-Aldrich) and total protein was determined by BCA (Thermo Fisher Scientific). Lysates or supernatants were separated by SDS-PAGE (NuPAGE, Thermo Fisher Scientific) and blotted onto nitrocellulose, nytrane membranes (GE healthcare, Chicago, IL, USA). Anti-mouse caspase-1, full-length and activated p20 fragment (mAb Casper-1, Adipogen Life Sciences, Liestal, Switzerland), ASC (anti-Asc, pAb (AL177), Adipogen Life Sciences), NLRP3 (mAB Cryo-2, Adipogen Life Sciences), IL-1 ß (anti-mIL-1β R&D Systems) were used as primary and horseradish-peroxidase-conjugated anti-rabbit and anti-mouse IgG (both Cell Signaling Technology, Beverly, MA, USA) as secondary antibodies. Chemiluminescent substrate (Biozym Scientific GmbH, Hessisch Oldendorf, Germany) was used for visualization.
Peukert K., Steinhagen F., Fox M., Feuerborn C., Schulz S., Seeliger B., Schuss P., Schneider M., Frede S., Sauer A., Putensen C., Latz E., Wilhelm C, & Bode C. (2022). Tetracycline ameliorates silica-induced pulmonary inflammation and fibrosis via inhibition of caspase-1. Respiratory Research, 23, 21.
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2

Liver Protein Expression Analysis

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Liver tissue and cells were lysed in RIPA buffer (R&D) supplemented by a protease inhibitor (Thermo Scientific) and a phosphatase inhibitor cocktail (Thermo Scientific). Protein concentrations were quantified using BCA Protein Assay Kit (Pierce). Equal amounts of protein were separated by SDS–PAGE and transferred onto nitrocellulose membranes. After blocking with 5% milk for 1 hour at room temperature, the membranes were incubated with primary antibodies (anti-mouse TREM2, anti-mouse GAPDH, anti-mouse Capase-3, anti-mouse Cleavage Capase-3, anti-mouse ADAM10, anti-mouse ADAM17, anti-mouse S1PR1, anti-mouse S1PR2, anti-mouse SPHK1, anti-mouse SPHK2, anti-mouse Akt, anti-mouse Phospho-Akt (Ser473), anti-human TREM2 and anti-human GAPDH), followed by incubation with the appropriate HRP-conjugated secondary antibodies and developed with ECL. Images were recorded using a ChemiDoc™ Touch Imaging System Surpasses (Bio-Rad).
Wang X., He Q., Zhou C., Xu Y., Liu D., Fujiwara N., Kubota N., Click A., Henderson P., Vancil J., Marquez C.A., Gunasekaran G., Schwartz M.E., Tabrizian P., Sarpel U., Fiel M.I., Diao Y., Sun B., Hoshida Y., Liang S, & Zhong Z. (2022). Prolonged hypernutrition impairs TREM2-dependent efferocytosis to license chronic liver inflammation and nonalcoholic steatohepatitis development. Immunity, 56(1), 58-77.e11.
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